Biochemical and Biophysical Research Communications 525 (2020) 334e340

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Biochemical and Biophysical Research Communications

journal homepage: www.elsevier.com/locate/ybbrc

Advanced glycation end products enhance M1 macrophage

polarization by activating the MAPK pathway
Sunyue He a, Qiuyue Hu a, Xiaoyuan Xu a, Yixin Niu a, Youming Chen c, Yao Lu a,
Qing Su a, **, Li Qin a, b, *
a
Department of Endocrinology, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, 1665 Kongjiang Road, Shanghai, 200092, China
b
Department of Endocrinology, Xinhua Hospital Chongming Branch, School of Medicine, Shanghai Jiaotong University, 25 Nanmen Road, Shanghai, 202150,
China
c
Department of Cardiology, Xinhua Hospital, School of Medicine, Jiao Tong University, 1665 Kongjiang Road, Shanghai, 200092, China

a r t i c l e i n f o a b s t r a c t

Article history: Background: b-cell dysfunction is one of the core pathogenetic mechanisms of type 2 diabetes mellitus
Received 19 November 2019 (T2DM). However, there are currently no effective therapeutic strategies to preserve b-cell mass and
Accepted 7 February 2020 function. The role of islet macrophage phenotype reprogramming in b-cell dysfunction has attracted
Available online 21 February 2020
great attention. Given that advanced glycation end products (AGEs) are major pathogenic factors in
T2DM, we investigated the effect of AGEs on macrophage activation and their role in b-cell dysfunction.
Keywords:
Methods: We examined cytokine secretion, M1 and M2 macrophage-associated marker expression and
AGEs
MAPK phosphorylation levels in AGEs-stimulated macrophages. MIN6 cells were cocultured with AGEs-
Macrophage polarization
MAPK
pretreated macrophages to study the effect of AGEs-induced macrophage activation on b-cell
b-Cell dysfunction dysfunction.
T2DM Results: We found that AGEs treatment significantly enhanced macrophage secretion of proinflammatory
cytokines. The expression of M1 macrophage markers, such as iNOS and the surface marker CD11c, was
significantly upregulated, whereas the expression of M2 macrophage markers, such as Arg1 and CD206,
was reciprocally downregulated upon AGEs stimulation. AGEs treatment predominantly activated the
MAPK pathway, and the inhibition of the MAPK pathway partially attenuated the AGEs-induced polar-
ization of macrophages. In addition, coculture with AGEs-pretreated macrophages significantly inhibited
the expression of molecules involved in b-cell function and was accompanied by the impairment of
glucose-stimulated insulin secretion (GSIS) in MIN6 cells.
Conclusion: AGEs enhance the expression of proinflammatory molecules by activating the MAPK
pathway. Moreover, these data imply that AGEs induce macrophage M1 phenotype polarization but
restrain M2 polarization, which might contribute to b-cell dysfunction in the pathogenesis of T2DM.
© 2020 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND
license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

Type 2 diabetes mellitus (T2DM) is mainly characterized by

Abbreviations: AGEs, advanced glycation end products; BSA, bovine serum al-
bumin; T2DM, type 2 diabetes mellitus; iNOS, inducible nitric oxide synthase; Arg1,
arginase 1; MAPK, mitogen-activated protein kinase; KLF4, Kruppel-like factor 4;
LPS, lipopolysaccharide; GSIS, glucose-stimulated insulin secretion.
* Corresponding author. Department of Endocrinology, Xinhua Hospital
Chongming Branch, School of Medicine, Shanghai Jiaotong University, 25 Nanmen
Road, Shanghai, 202150, China.
** Corresponding author. Department of Endocrinology, Xinhua Hospital, School
of Medicine, Shanghai Jiaotong University, 1665 Kongjiang Road, Shanghai, 200092,
China.
E-mail addresses: suqingxinhua@163.com (Q. Su), qinli@xinhuamed.com.cn
(L. Qin).
hyperglycemia, insulin resistance and impaired insulin secretion
[1]. It is becoming an expanding global health problem. Recently,
the onset of T2DM has been found in patients of a relatively young
age [2,3]. b-cell dysfunction is one of the core pathogenetic
mechanisms of T2DM and is also associated with poor glycemic
control and treatment failure. b-cell function starts to decline even
before the onset of impaired glucose tolerance [4]. Therefore, it is of
value to maintain or restore b-cell function in the treatment of
T2DM. However, there are no currently effective therapeutic stra-
tegies to preserve b cell mass and function.

https://doi.org/10.1016/j.bbrc.2020.02.053
0006-291X/© 2020 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
 S. He et al. / Biochemical and Biophysical Research Communications 525 (2020) 334e340
Macrophages are a heterogeneous population of immune cells
that display remarkable plasticity and change their physiology in
response to various microenvironmental stimuli [5]. These changes
can produce different populations of cells with distinct functions:
the classically activated macrophages (or M1) are induced by
interferon-g (IFN-g) and/or lipopolysaccharide (LPS) whereas the
alternatively activated macrophages (or M2) are induced by inter-
leukin (IL)-4 and/or IL-13 [6]. M1 polarized macrophages are
characterized by high expression levels of inducible nitric oxide
synthase (iNOS), the ability to produce large amounts of proin-
flammatory cytokines, such as IL-1b, IL-6, and TNF-a and increased
levels of NO. In contrast, M2 polarized macrophages are charac-
terized by increased expression level of CD206, CD163, arginase 1
(Arg1), and Kruppel-like factor 4 (KLF4) and the secretion of anti-
inflammatory cytokines [7].
It has long been known that macrophages are present within
islets [8] and play a crucial role in maintaining b-cell function and
proliferation [9]. However, several studies have found an increased
number of islet macrophages in rodent T2DM models [10,11] as
well as human T2DM patients [12] with almost all macrophages
polarized toward the M1-like phenotype [10]. This shift has been
shown to contribute to b-cell dysfunction in T2DM mouse models
[13]. In addition, the IL-1 pathway has been shown to play a key role
in T2DM and macrovascular complications in preclinical studies
[14]. A growing body of evidence suggests that macrophages are
the main source of proinflammatory cytokines within islets [9,13].
Islet macrophages cause b-cell dysfunction by secreting IL-1b and
have been widely studied [13,14]. It has also been indicated that
inhibiting the IL-1 pathway can restore b-cell function and alleviate
hyperglycemia in both humans and rodents [15e18]. Thus, the
phenotypic reprograming of islet macrophages plays a causative
role in the development of b-cell dysfunction.
Advanced glycation end products (AGEs) are a complex and
heterogeneous group of compounds produced by nonenzymatic
glycation and oxidation of proteins, nucleic acids, and lipids [19].
AGEs are known to accumulate in the plasma and tissues, such as
islets, in diabetic patients and experimental models [20,21]. In
recent years, the effect of islet inflammation and AGEs-induced islet
injury in b-cell failure have attracted great attention. However, its
molecular mechanisms have not been fully elucidated. Previous
studies from our group and others suggest that AGEs may increase
ROS production and impair mitochondrial function, giving rise to
dysfunctional b-cell insulin secretion [22e24]. In addition,
administration of AGEs in C57BL/6J mice not only caused insulin
secretion deficiency and enhanced insulitis score, but also signifi-
cantly increased islet macrophages infiltration [25]. Nonetheless,
which subset of macrophages increased in islet upon AGEs treat-
ment is still unclear. And it still remains to be explored whether
AGEs induce b-cell damage by regulating the polarization of islet
macrophages. Therefore, the present study aimed to test whether
AGEs enhance macrophage M1 polarization as a potential mecha-
nism underlying b-cell dysfunction.
2. Materials and methods
2.1. Antibodies
iNOS (Sigma, N7782), NLRP3 (Adipogen, clone Cryo-2), Arg1
(Abcam, ab124917), CD206 (Abcam, ab125028), KLF4 (Abcam,
ab214666), a-tubulin (Abcam, ab7291), phospho-c-Jun NH2-ter-
minal kinase (p-JNK; Cell Signaling Technology, 4671s), JNK (Cell
Signaling Technology, 9252s), phospho-p38 MAPK (Cell Signaling
Technology, 4511), p38 MAPK (Abclonal, AP0526), phospho-
extracellular signal-regulated protein kinases 1 and 2 (p-ERK1/2;
Millipore, 05e797F), ERK1/2 (Millipore, 06e182), Pdx1(Cell
335
Signaling Technology, D59H3), Insulin(Cell Signaling Technology,
C27C9).
2.2. Cell culture
Murine Raw 264.7 cells were cultured in DMEM containing
25 mM glucose supplemented with 10% heat-inactivated FBS.
Murine MIN6 b-cells were cultured in RPMI 1640 supplemented
with 10% heat-inactivated FBS and 50 mM b-mercaptoethanol. All
culture dishes were supplemented with 1% penicillin/streptomycin
and maintained at 37  C containing 5% CO2. To investigate the effect
of AGEs on macrophage polarization, Raw 264.7 cells were treated
with different concentrations of AGEs or BSA (200 mg/L) as a
negative control for 24 h. To analyze the effect of AGEs-pretreated
macrophages on b-cell function, Raw 264.7 cells were plated in
the Boyden chamber insert containing 0.4 mm porous membranes
(polyethylene terephthalate membrane, Corning) and treated with
AGEs or BSA for 24 h followed by coculture with MIN6 cells for
another 24 h.
2.3. Western blotting analysis
Western blotting analysis was performed as described previ-
ously [25]. The following secondary antibodies were used: horse-
radish peroxidase-coupled secondary anti-rabbit or anti-mouse
antibodies. Signals were visualized using a Gel Imaging System
(Tanon). The band densitometry was quantified with ImageJ
software.
2.4. Proinflammatory cytokine detection
The concentrations of IL-1-b, IL-6 and TNF-a in cell-free culture
supernatants were measured using enzyme-linked immunosor-
bent assay (ELISA) kits (Shanghai Westang Biotech Inc., China) by
following the manufacturer’s protocol.
2.5. Nitrite generation assay
The nitrite level in the culture supernatant was measured by a
NO Assay kit (CibZist Fan Co.). The absorbance of each well was
measured at 540 nm with a microplate reader (BioTek, Germany),
and the actual concentration was calculated according to the
manufacturer’s instructions.
2.6. Flow cytometry
F4/80 was used as a pan-macrophage marker, and CD11c was
used as an M1 macrophage marker. To detect the cell-surface
expression of CD11c, Raw 264.7 cells were cultured with BSA,
AGEs or LPS for 24 h. After being washed, the cells from different
treatment groups were incubated with APC-conjugated anti-mouse
F4/80 and PE-conjugated anti-mouse CD11c antibodies in PBS for
30 min at 4  C and then washed twice with PBS. Cell sorting was
performed using a CytExpert flow cytometry system and analyzed
by FlowJo software (TreeStar, Ashland, OR). The mean fluorescence
intensity was used for the quantitative analysis.
2.7. Glucose-stimulated insulin secretion (GSIS)
MIN6 cells were cultured in 12-well plates and cocultured with
BSA or AGEs-pretreated Raw 264.7 macrophages for 24 h. Prior to
the experiment, the MIN6 cells were washed three times with PBS
and then preincubated in Krebs-Ringer buffer (KRB; 129 mM NaCl,
4.8 mM KCl, 2.5 mM CaCl2, 5 mM NaHCO3, 1.2 mM KH2PO4, 1.2 mM
MgSO4, 10 mM HEPES and 2 mg/ml BSA) containing 2.8 mM glucose
336 S. He et al. / Biochemical and Biophysical Research Communications 525 (2020) 334e340
for 1 h. Subsequently, the cells were incubated with KRB buffer
containing either 2.8 mM glucose or 22.4 mM glucose for another
1 h at 37  C in a humidified 5% CO2 atmosphere. This glucose
stimulation was performed in triplicate for each passage. The su-
pernatant was collected and centrifuged to remove any cells or
debris, and the insulin concentration was measured using an ELISA
kit (Shanghai Westang Biotech Inc., China) in accordance with the
manufacturer’s instructions.
2.8. Immunofluorescence
MIN6 cells were cultured in 6-well plates and then cocultured
with BSA or AGEs-pretreated Raw 264.7 macrophages for 24 h.
MIN6 cells were fixed with 4% paraformaldehyde for 20 min at RT
and washed three times with PBS for 5 min per wash. After
washing, the cells were blocked in 1% BSA at RT and then stained
primary antibodies against Pdx1 or Insulin in 1% BSA at 4  C over-
night. The cells were then washed three times with 5% PBS-Tween
(PBST) for 5 min per wash and stained with an Alexa Fluor-488 or
Fluor-549 secondary antibody (1:1000, dilution; Abcam, Cam-
bridge, UK) for 1 h at RT. After desiccation, the cells were coun-
terstained with DAPI (Sigma, St. Louis, MO). All cells were visualized
with a laser confocal scanning microscope.
2.9. Statistical analysis
All data were shown as the mean ± standard deviation (SD). Two
groups were compared by unpaired student’s t-test. Comparison
between multiple groups was done by one-way ANOVA test. A P
value < 0.05 was considered statistically significant. Statistical
calculations were performed using GraphPad PRISM 6.0 statistical
software.
3. Results
3.1. Effect of AGEs on raw 264.7 macrophage viability and
morphology
The treatment of cells with AGEs did not affect the viability at
24 h (Fig. S2A). Therefore, 200 mg/L AGEs were used for the sub-
sequent experiments. LPS is known to induce monocyte/macro-
phage polarization to the M1 phenotype; therefore, it was used as a
positive control in our studies. The hematoxylin and eosin staining
Fig. 1. Effect of AGEs on the expression of proinflammatory cytokines and M1/M2-related marker in macrophages. (a) The levels of the cytokines IL-1b, IL-6, and TNF-a were
determined by ELISA. (b) Protein levels of iNOS, NLRP3, Arg1, CD206, KLF4 and a-tubulin were detected by western blotting. (c) The macrophage M1 surface antigen CD11c was
analyzed by flow cytometry. (d) NO generation. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.
showed that macrophages incubated with BSA were almost
spherical and fusiform; however, the surface of the macrophages
incubated with AGEs and LPS had increased numbers of “synapses”.
Similarly, the morphological characteristics of the AGEs-treated
cells were similar to those of cells in the M1-positive groups
(Fig. S2B).
3.2. AGEs induce proinflammatory cytokine expression and
promote M1 and inhibit M2 macrophage polarization
To assess whether AGEs mediate M1 macrophage polarization in
Raw 264.7 cells, we examined the expression of M1-and M2-
associated markers. AGEs treatment upregulated the expression
of proinflammatory cytokines, such as IL-1b, TNF-a, and IL-
6(Fig. 1B, Figure S3A). Furthermore, AGEs treatment increased the
expression of M1-related markers, such as NOS2, NLRP3, and
SOCS3, but inhibited the expression of M2-related markers, such as
Arg1, Mrc1, and KLF4 gene transcription (Figure S3B, C). Accord-
ingly, western blot analysis also revealed that AGEs treatment
obviously upregulated iNOS and NLRP3 but downregulated Arg1,
CD206 and KLF4 protein expression in a dose-dependent manner
(Fig. 1A). Flow cytometry analyses demonstrated that AGEs treat-
ment markedly increased the expression of the M1 surface marker
CD11c (Fig. 1C). Moreover, the NO generation of the AGEs-treated
group was significantly higher than that of the BSA-treated group
(Fig. 1D). In summary, these data supported the hypothesis that
AGEs skew macrophage polarization toward a proinflammatory
phenotype.
3.3. AGEs activate MAPK pathways in macrophages
To elucidate the molecular and cellular pathways by which AGEs
regulate macrophage activation, western blotting was used to
examine the activation of the MAPKs (JNK, ERK1/2 and p38) in
macrophages. As shown in Fig. 2A and B, AGEs treatment signifi-
cantly enhanced the phosphorylation of MAPK family members
(JNK, ERK1/2, and p38) in a time- and dose-dependent manner. The
phosphorylation of all three MAPKs was detectable as early as
15 min, and peaked at 30 min, followed by a slight drop after
exposure to AGEs. These data indicate that activation of the MAPK
signaling pathway is a possible mechanism to explain the proin-
flammatory effects of AGEs.
 S. He et al. / Biochemical and Biophysical Research Communications 525 (2020) 334e340 337

Fig. 2. The role of the MAPK pathway in AGE-induced proinflammatory cytokine and M1/M2-related marker expression in macrophages. (a, b) Western blot analysis of the
phosphorylated and total levels of JNK, ERK and p38 and analyzed based on Densitometric statistics. (c) The levels of the cytokines IL-1b, IL-6, and TNF-a were determined by ELISA.
(d) Protein levels of iNOS, NLRP3, KLF4 and a-tubulin were detected by western blotting. (e) The macrophage M1 surface antigen CD11c was analyzed by flow cytometry. (f) NO
generation. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

3.4. AGEs-induced M1 macrophage polarization is mediated by
MAPK pathways
We next determined the role of these kinases in AGEs-induced
M1 macrophage polarization. Raw 264.7 cells were pretreated
with a JNK inhibitor (SP600125, 10 mM), ERK1/2 inhibitor (U0126,
10 mM), or p38 MAPK inhibitor (SB203580, 10 mM) for 1 h. After
being treated with the inhibitors, the cells were incubated with
AGEs for 8 h for mRNA analyses or 24 h for M1-and M2-related
marker detection. As shown in Fig. 2CeF, Figure S4, JNK, ERK1/2
and p38 MAPK inhibitor treatment significantly decreased the
AGEs-induced production of NO and proinflammatory cytokines
(IL-1b and IL-6). AGEs-induced alterations in KLF4, iNOS, TNF-a,
NLRP3, and Mrc1 expression were prominently attenuated by the
inhibition of JNK and ERK1/2. Furthermore, ERK1/2 inhibitor
(U0126) treatment markedly inhibited the AGEs-induced upregu-
lation of the expression of the M1 surface marker CD11c. However,
p38 MAPK inhibitor pretreatment resulted in an unexpected
338 S. He et al. / Biochemical and Biophysical Research Communications 525 (2020) 334e340

increase in TNF-a secretion. In conclusion, these data indicate that
MAPK activation is required for AGEs-induced M1 macrophage
polarization.
3.5. AGEs induce b-cell dysfunction via macrophage activation
To further investigate whether AGEs induce b-cell dysfunction
by regulating macrophage polarization, we used a coculture sys-
tem. In MIN6 cells cocultured with AGEs-pretreated macrophages,
the expression of Pdx1, Insulin and MAFA was significantly reduced
compared to that in cells cocultured with BSA-pretreated macro-
phages (Fig. 3C, E, F). In addition, coculture with AGEs-treated
macrophages also impaired GSIS in MIN6 cells (Fig. 3B). These re-
sults suggest that AGEs-pretreated macrophages are involved in b-
cell dysfunction. Interestingly, the expression of CCL2/MCP-1 was
also markedly enhanced MIN6 cells cocultured with AGEs-
pretreated macrophages compared to those cocultured with BSA-
pretreated macrophages (Fig. 3D).
4. Discussion
In the present study, we found that AGEs treatment polarized
macrophages to the proinflammatory M1 phenotype and inhibited
M2 polarization, which was partially linked to enhanced

Fig. 3. Effect of AGE-treated macrophages on b-cell function. (a) Study design. (b) GSIS. (c) Relative gene expression of Pdx1, Insulin, MAFA, and CCL2. (c, d). Expression level of
Pdx1 and Insulin detected by immunofluorescence. **P < 0.01 and ***P < 0.001.
 S. He et al. / Biochemical and Biophysical Research Communications 525 (2020) 334e340
intracellular MAPK signaling. Moreover, our work shows for the
first time that AGEs play a role in the development of b-cell
dysfunction by regulating macrophage phenotype reprogramming
(see Fig. 4).
LPS is known to polarize monocytes/macrophages to the M1
phenotype. Our data confirm the effects of LPS on the upregulation
of M1 macrophage marker expression in Raw 264.7 cells. Similarly,
AGEs treatment significantly upregulated the expression of IL-1b,
IL-6, TNF-a, SOCS3, NLRP3 and CD11c, which are associated with
M1 macrophages, whereas AGEs treatment downregulated the
expression of the M2-related marker CD206. These results are
consistent with those of previous studies showing that AGEs
polarize macrophages toward the proinflammatory state by
inducing the expression of proinflammatory molecules [26,27].
Differential NO metabolism is a well-defined and reliable
marker to discriminate the two activation states of macrophages.
M1 macrophages express high levels of iNOS, which is involved in
the synthesis of NO from arginine. However, M2 macrophages have
increased levels of Arg1, which competes for the arginine substrate,
thereby reducing NO production [28]. In this experiment, we
observed that AGEs treatment induced a significant increase in
iNOS expression and modestly inhibited Arg1 expression in Raw
264.7 macrophages, and led to increased NO production. However,
Jin et al. found that AGEs predominantly enhance iNOS expression
without affecting Arg1 expression in BMDMs [26]. These discrep-
ancies may be related to differences in the cell lines and experi-
mental conditions.
Recent studies have also found that KLF4 acts as a transcrip-
tional regulator of macrophage differentiation, which is the subject
of intensive research. KLF4 is expressed in a stage-specific pattern
during myelopoiesis and promotes monocyte/macrophage polari-
zation. Its expression is robustly induced in M2 macrophages but
strongly reduced in M1 macrophages [29]. Our results demonstrate
that KLF4 mRNA abundance and protein levels are strongly sup-
pressed by AGEs treatment compared with BSA treatment. More-
over, consistent with previous experiments, IL-4 stimulation
dramatically promoted KLF4 gene expression, yet the induction of
KLF4 was manifestly decreased in cells pretreated with AGEs.
Collectively, these results show that AGEs treatment increases the
expression of proinflammatory mediators and M1 surface markers
and suppresses the production of critical phenotype regulators in
macrophages. These data indicate that AGEs induce M1
Fig. 4. Schematic illustration of the effect of AGEs on macrophage activation and the underlying mechanism. AGE treatment enhances the expression of proinflammatory
molecules through the activation of the MAPK signaling pathway. Moreover, AGEs induce macrophage M1 phenotype polarization but restrain M2 polarization, which might
contribute to b-cell dysfunction in the pathogenesis of T2DM.
339
macrophage phenotype polarization as well as inhibit M2 polari-
zation and reduce IL-4-induced M2 skewing of macrophages.
Since macrophage polarization requires the activation of specific
transcription factors, the possible pathways involved in AGEs-
induced M1 macrophage polarization could be further elucidated.
Here, we observed that AGEs pretreatment of macrophages mark-
edly increased the phosphorylation of MAPK signaling pathway
proteins in a time- and dose-dependent manner. In the inflam-
matory response, MAPKs have critical effects on the regulation of
proinflammatory cytokines and mediators [30]. Several studies
have reported that the increased phosphorylation of members of
the MAPK family is involved in macrophage activation triggered by
various stimuli [31e33]. Additionally, Liu et al. demonstrated that
AGEs induce inflammatory responses and the release of inflam-
matory mediators through the MAPK signaling pathway in vascular
adventitial fibroblasts [34]. These results further support our re-
sults showing that AGEs may activate macrophages via the MAPK
signaling pathway. The inhibition of MAPK signaling pathway
activation by various inhibitors significantly decreased the pro-
duction of NO and M1-related proinflammatory molecules. How-
ever, pretreatment with a p38 MAPK inhibitor resulted in an
unexpected increase in TNF-a secretion, which may be related to
the complex regulation between intracellular signaling pathways.
In addition to promoting inflammatory responses, p38 has complex
effects on inflammation regulation, including activating feedback
pathways that downregulate inflammation in innate immunity
[35]. In summary, these data suggest that the AGEs-induced po-
larization of macrophages to the proinflammatory phenotype
might at least partially be linked to enhanced intracellular MAPK
signaling.
As mentioned above, b-cell dysfunction, one of the core path-
ogenetic mechanisms of T2DM, is tightly associated with islet
macrophage phenotype reprogramming [36]. Our data show that
AGEs induce unfavorable M1/M2 macrophage polarization in vitro.
Furthermore, we observed that in MIN6 cells cocultured with AGEs-
pretreated macrophages, b-cell function was significantly lower
than that in MIN6 cells cocultured with BSA-pretreated macro-
phages and was accompanied by impaired GSIS. This result sug-
gests that AGEs-induced macrophage activation may be involved in
the pathological development of b-cell dysfunction. In addition,
AGEs-pretreatment of Raw 264.7 cells induced CCL2/MCP-1
expression in MIN6 cells. These results imply that communication
340 S. He et al. / Biochemical and Biophysical Research Communications 525 (2020) 334e340
via proinflammatory cytokines and chemokines between M1-like
macrophages and b-cells forms a vicious cycle that further propa-
gates and perpetuates inflammatory processes within islets, lead-
ing to b-cell dysfunction. Thus, our data prove that AGEs exposure
induces macrophage phenotype reprogramming, and plays an
important role in b-cell dysfunction. This study reveals a novel
pathological role and potential mechanisms by which AGEs affect
the pathogenesis of T2DM. Therefore, the mechanism of b-cell
dysfunction induced by AGEs-pretreated macrophages needs to be
further elucidated.
Author contributions
Sunyue He performed the experiments, analyzed the data and
wrote the manuscript; Qiuyue Hu contributed to the pathway
analysis and discussed the manuscript; Xiaoyuan Xu, Yixin niu and
Youming Chen contributed to the data collection and analysis; Yao
Lu contributed to the data interpretation and discussed the
manuscript; Qing Su and Li Qin designed and supervised study, as
well as revised manuscript.
Declaration of competing interests
The authors declare that they have no conflict of interest.
Acknowledgements
This study was supported by National Natural Science Founda-
tion of China (81670743, 81970669), Shanghai Municipal Health
Commission (201740173) and Shanghai Sailing Program
(18YF1415800).
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.bbrc.2020.02.053.
Transparency document
Transparency document related to this article can be found
online at https://doi.org/10.1016/j.bbrc.2020.02.053.
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